Compositions and methods for synthesizing 5'-Capped RNAs
Provided herein are methods and compositions for synthesizing 5′Capped RNAs wherein the initiating capped oligonucleotide primers have the general form m7 Gppp[N 2′Ome ] n [N] m wherein m7 G is N7-methylated guanosine or any guanosine analog, N is any natural, modified or unnatural nucleoside, “n” can be any integer from 0 to 4 and “m” can be an integer from 1 to 9.
1. A compound of the following formula:
wherein:
B 1 is adenine or N6-methyladenine;
B 10 is guanine;
R 1 is H or methyl;
R 2 is H, OH, alkyl, O-alkyl, halogen, a linker or a detectable marker;
R 3 is O-alkyl; and
R 4 is H, OH, O-methyl, or a detectable marker, or
a stereoisomer thereof,
wherein the compound is a salt of the formula.
2. The compound of claim 1 , wherein B 1 is adenine.
3. The compound of claim 1 , wherein B 1 is N6-methyladenine.
4. The compound of claim 1 , wherein R 1 is H.
5. The compound of claim 1 , wherein R 2 is OH or O-methyl.
6. The compound of claim 1 , wherein R 3 is O-methyl.
7. The compound of claim 1 , wherein R 4 is O-methyl.
8. A compound of the following structure:
or a stereoisomer thereof, wherein the compound is a salt of the structure.
9. A compound of the following structure:
or a stereoisomer thereof, wherein the compound is a salt of the structure.
10. The compound of claim 1 , wherein the salt is a sodium salt, a lithium salt, or a potassium salt.
11. An RNA molecule comprising the compound or stereoisomer thereof according to claim 1 .
12. An isolated cell comprising an RNA molecule according to claim 11 .
13. A modified cell comprising a protein or a peptide translated from an RNA molecule according to claim 11 .
14. A pharmaceutical composition comprising an RNA molecule according to claim 11 and a pharmaceutically acceptable carrier.
15. An initiating capped oligonucleotide primer selected from the group consisting of:
a salt of m7 G 3′Ome pppA 2′Ome pG;
a salt of m7 G 3′Ome ppp(N6-methyladenine) 2′Ome pG;
a salt of m7 G 3′Ome pppApG; and
a salt of m7 G 3′Ome ppp(N6-methyladenine)pG.
16. An RNA molecule comprising the initiating capped oligonucleotide primer according to claim 15 .
17. An isolated cell comprising an RNA molecule according to claim 16 .
18. A modified cell comprising a protein or a peptide translated from an RNA molecule according to claim 16 .
19. A pharmaceutical composition comprising an RNA molecule according to claim 16 and a pharmaceutically acceptable carrier.
20. The compound of claim 8 , wherein the salt is a sodium salt, a lithium salt, or a potassium salt.
21. The compound of claim 9 , wherein the salt is a sodium salt, a lithium salt, or a potassium salt.
22. The initiating capped oligonucleotide primer of claim 15 , comprising a salt of m 7 G 3′Om epppA 2′Ome pG.
23. An RNA molecule comprising the initiating capped oligonucleotide primer according to claim 22 .
24. The initiating capped oligonucleotide primer of claim 15 , comprising a salt of m 7 G 3′Om eppp (N6-methyladenine) 2′Ome pg.
25. An RNA molecule comprising the initiating capped oligonucleotide primer according to claim 24 .
26. A modified cell comprising a protein or a peptide translated from an RNA molecule according to claim 23 .
27. A pharmaceutical composition comprising a cell comprising an RNA molecule according to claim 11 .
28. A pharmaceutical composition comprising a cell comprising an RNA molecule according to claim 16 .
29. The compound of claim 2 , wherein B 10 is guanine.
30. The compound of claim 3 , wherein B 10 is guanine.